Platelet Storage and Reduced Bacterial Proliferation In Platelet Products Using A Sialidase Inhibitor

Abstract

The present invention relates to methods and compositions for reducing sialidase activity and inhibiting bacterial proliferation of one or more bacteria in a platelet product preparation from one or more donors. In general, the method includes contacting the platelet product preparation with an amount of a sialidase inhibitor, to thereby obtain a sialidase inhibitor-treated platelet product preparation. Sialidase activity is reduced and the proliferation of one or more bacteria is inhibited, as compared to a platelet product preparation not subjected to the sialidase inhibitor treatment.

What is claimed is: 
     
         1 ) A method of increasing the storage time of a population of platelets wherein the platelets have a platelet surface and sialic acid loss is caused by the presence of active endogenous sialidase that has migrated to the platelet surface during storage, the method consists essentially of:
 a) obtaining a population of platelets from one or more individuals by separating platelets from blood of one or more individuals;   b) treating the population of platelets with a platelet additive solution (PAS) that comprises an amount ranging between about 0.1 mM and about 100 mM of one or more sialidase inhibitors and one or more PAS components, wherein said treating is performed by adding the population of platelets to the PAS, by adding PAS to the population of platelets, or by both, to thereby obtain a platelet composition of treated platelets; and   c) storing the treated platelets at a temperature ranging between about 1° C. and about 6° C. for a period of time ranging between about 1 day and 30 days, wherein cleavage by the endogenous sialidase of sialic acid and sialic acid loss on the platelet surface is reduced, as compared to isolated platelets not subjected to Step b); and wherein upon transfusion of the platelet composition into a recipient, circulation time of platelets is increased by at least 10% and platelet clearance of the platelets is reduced, as compared to circulation time and platelet clearance of platelets that have not been subjected to Step b)   wherein the one or more sialidase inhibitors is selected from the group consisting of: fetuin; 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA); ethyl (3R,4R, 5 S)-5-amino-4-acetamido-3-(pentan-3-yloxy)-cyclohex-1-ene-1-carboxylate); (2R,3R,4S)-4-guanidino-3-(prop-1-en-2-ylamino)-2-((1R,2R)-1,2,3-trihydroxypropyl)-3,4-dihydro-2H-pyran-6-carboxylic acid; (4S,5R,6R)-5-acetamido-4-carbamimidamido-6-[(1R,2R)-3-hydroxy-2-methoxypropyl]-5,6-dihydro-4H-pyran-2-carboxylic acid; (1S,2S,3S,4R)-3-[(1S)-1-acetamido-2-ethyl-butyl]-4-(diaminomethylideneamino)-2-hydroxy-cyclopentane-1-carboxylic acid; and a pharmaceutically acceptable salt thereof.   
     
     
         2 ) The method of  claim 1 , further including treating the population of platelets with the PAS within a time frame, wherein the time frame is in a range between about 1 minute to about 8 hours. 
     
     
         3 ) The method of  claim 1 , wherein the PAS components includes a salt, a phosphate source, a citrate source, a carbon source, an acetate source, or any combination thereof. 
     
     
         4 ) The method of  claim 1 , wherein the PAS is maintained at a pH ranging between about 6.4 and about 7.6. 
     
     
         5 ) The method of  claim 3 , wherein the phosphate source is present in an amount ranging from about 5 mM to about 50 mM, and wherein the phosphate source is selected from the group consisting of sodium monophosphate, sodium diphosphate, sodium triphosphate, and a combination thereof. 
     
     
         6 ) The method of  claim 3 , wherein the citrate source is present in an amount ranging from about 2mM to about 20mM, and wherein the citrate source is selected from the group consisting of monosodium citrate, disodium citrate, trisodium citrate, citric acid, and a combination thereof. 
     
     
         7 ) The method of  claim 3 , wherein the carbon source is present in an amount ranging from about 0.5 mM to about 50 mM, and wherein the carbon source is selected from the group consisting of acetate, glucose, and sucrose. 
     
     
         8 ) The method of  claim 3 , wherein the acetate source is present in an amount ranging from about 10 mM to about 50 mM, and wherein the acetate source is selected from the group consisting of sodium acetate, potassium acetate, magnesium acetate, and a combination thereof. 
     
     
         9 ) The method of  claim 3 , wherein the salt is selected from the group consisting of a sodium source, a chloride source, a potassium source, a magnesium source, a calcium source, and a combination thereof. 
     
     
         10 ) The method of  claim 9 , wherein the sodium source is selected from the group consisting of sodium chloride, sodium citrate, sodium acetate, sodium phosphate, and a combination thereof. 
     
     
         11 ) The method of  claim 9 , wherein the chloride source is selected from the group consisting of sodium chloride, magnesium chloride, potassium chloride, and a combination thereof. 
     
     
         12 ) The method of  claim 9 , wherein the potassium source is selected from the group consisting of potassium chloride, potassium citrate, potassium acetate, potassium phosphate, potassium sulfate, and a combination thereof. 
     
     
         13 ) The method of  claim 9 , wherein the magnesium source is selected from the group consisting of magnesium chloride, magnesium citrate, magnesium sulfate, and a combination thereof. 
     
     
         14 ) The method of  claim 9 , wherein the calcium source is selected from the group consisting of calcium chloride, calcium acetate, calcium citrate, and a combination thereof. 
     
     
         15 ) The method of  claim 1 , further comprising rewarming the platelet composition to room temperature prior to transfusion to the recipient. 
     
     
         16 ) The method of  claim 1 , wherein step b) further comprises adding the population of platelets to one or more glycan-modifying agents, adding one or more glycan-modifying agents to the population of platelets, or both, wherein the one or more glycan-modifying agents comprise CMP-sialic acid or a CMP-sialic acid precursor. 
     
     
         17 ) The method of  claim 1 , wherein step b) further comprises adding the population of platelets to an enzyme, adding an enzyme to the population of platelets, or both that converts the CMP-sialic acid precursor to CMP-sialic acid. 
     
     
         18 ) The method of  claim 1 , wherein step b) further comprises adding the population of platelets to one or more glycan-modifying agents, adding one or more glycan-modifying agents to the population of platelets, or both, wherein the one or more glycan-modifying agents comprise UDP-galactose. 
     
     
         19 ) The method of  claim 1 , wherein step b) further comprises adding the population of platelets to one or both of two glycan-modifying agents, adding one or both of two glycan-modifying agents to the population of platelets, or both, wherein the two glycan-modifying agents are CMP-sialic acid and UDP-galactose. 
     
     
         20 ) The method of  claim 1 , wherein the method further inhibits proliferation of one or more bacteria found in a platelet composition. 
     
     
         21 ) The method of  claim 20 , wherein the one or more bacteria are selected from the group consisting of:  Aspergillus, Bacillus  sp.,  Bacteroides eggerthii, Candida albicans, Citrobacter  sp.,  Clostridium perfringens, Corynebacterium  sp.,  Diphtheroid, Enterobacter aerogenes, Enterobacter amnigenus, Enterobacter cloacae, Enterococcus avium, Enterococcus faecalis, Escherichia coli, Fusobacterium  spp.,  Granulicatella adiacens, Heliobacter pylori, Klebsiella  sp.,  Klebsiella pneumonia, Klebsiella oxytoca, Lactobacillus  sp.,  Listeria  sp.,  Micrococcus  sp.,  Peptostreptococcus, Proteus vulgaris, Pseudomonas  sp.,  Pseudomys oxalis, Propionibacterium  sp.,  Salmonella  sp.,  Serratia  sp.,  Serratia marcescens, Staphylococcus  sp., Coagulase-negative  Staphylococcus, Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus  sp.,  Streptococcus gallolyticus, Streptococcus bovis, Streptococcus pyogenes, Streptococcus viridans,  and  Yersinia enterocolitica.    
     
     
         22 ) The method of  claim 20 , further comprising a step of assessing a proliferation level in the sialidase inhibitor-treated platelet product preparation, to thereby obtain a treated proliferation level, and comparing the treated proliferation level to an untreated proliferation level, wherein the untreated proliferation level is a proliferation level in a platelet product preparation not treated with the sialidase inhibitor, and wherein the treated proliferation level is less than the untreated proliferation level.